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KMID : 0613820100200010097
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Journal of Life Science
2010 Volume.20 No. 1 p.97 ~ p.102
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Transconjugation for Molecular Genetic Study of Streptomyces platensis Producing Transglutaminase
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Bae Se-Jung
Jo Yang-Ho
Choi Sun-Uk
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Abstract
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Streptomyces platensis YK-2, newly isolated from forest soil, produces transglutaminase (TGase), which catalyses an acyl transfer reaction between the primary grade amine and protein or γ-carboxyamide group of peptide bound glutamine residues. For a molecular genetic study of S. platensis, an effective transformation method was established by using a conjugal transfer of DNA from Escherichia coli to spores of actinomycetes. The highest transconjugation frequency of S. platensis was obtained on an MS medium containing 50 mM MgCl₂, using 5×10? E. coli as a DNA donor and 1×10? spores without heat treatment as a host. We also identified that S. platensis contains a single attB site within an ORF encoding a pirin-homolog, and that its attB site sequence shows high homology to that of S. logisporoflavus. In addition, it was confirmed by phenotypic analyses of exconjugants that the introduction of heterologous DNA into the attB site of the S. platensis chromosome does not affect its morphological differentiation and TGase production.
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KEYWORD
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Streptomyces platensis YK-2, transformation, attB site, transglutaminase
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